CN107022528A - A kind of culture medium and its application for being used to cultivate hybridoma - Google Patents
A kind of culture medium and its application for being used to cultivate hybridoma Download PDFInfo
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- CN107022528A CN107022528A CN201710354795.XA CN201710354795A CN107022528A CN 107022528 A CN107022528 A CN 107022528A CN 201710354795 A CN201710354795 A CN 201710354795A CN 107022528 A CN107022528 A CN 107022528A
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- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/10—Cells modified by introduction of foreign genetic material
- C12N5/12—Fused cells, e.g. hybridomas
- C12N5/16—Animal cells
- C12N5/163—Animal cells one of the fusion partners being a B or a T lymphocyte
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- C12N2500/00—Specific components of cell culture medium
- C12N2500/05—Inorganic components
- C12N2500/10—Metals; Metal chelators
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- C12N2500/22—Zinc; Zn chelators
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- C12N2500/00—Specific components of cell culture medium
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- C12N2500/00—Specific components of cell culture medium
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- C12N2500/46—Amines, e.g. putrescine
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Abstract
The present invention relates to a kind of culture medium for hybridoma and its application, the culture medium contains following components on the basis of basal medium：P-methylbenzoic acid, lipoic acid, linoleic acid, putrescine, monoethanolamine, zinc sulfate, glutathione；Further relate to a kind of method for cultivating hybridoma and thus obtain Hybridoma Cell Culture thing.The Growth of Hybridoma Cell rule of the medium culture of the present invention, compared with traditional Hybridoma Cell Culture base, the hybridoma that culture is obtained in same time under the culture medium equal conditions of the present invention is more, cell volume is bigger, cellular morphology is fuller, cell viability is higher, and the duration cultivated is longer, can be preferably applied to the production of monoclonal antibody.
Description
Technical field
The present invention relates to field of cell culture, more specifically it relates to a kind of culture medium for being used to cultivate hybridoma and
It is applied, and is further related to a kind of method for cultivating hybridoma and is thus obtained Hybridoma Cell Culture thing.
Background technology
Monoclonal antibody with the specificity of its height and sensitivity, immunology detection, the affinity purification of destination protein and
Therapeutic field of tumor is used widely.Hybridoma technology is produced particularly important one during monoclonal antibody
Individual link, still, monoclonal antibody is prepared with the technology, its experiment link is more, and the cycle is long, influence factor is complicated, is subject to
Setback.Early stage especially is produced in hybridoma, because emerging hybridoma viability is poor, its growing state is easily because of environment
Change and change, it is impossible to be well adapted for vitro tissue condition of culture, it is easy to die young.Existing Hybridoma Cell Culture base example
Such as IMDM culture mediums are during culture hybridoma, and cell growth is unstable, and growth rate is not high, and culture medium is carried to cell
The duration of energy supply amount is not long.
Accordingly, it would be desirable to which a kind of new culture medium is used to cultivate hybridoma.
The content of the invention
To solve problem above, the invention provides a kind of culture medium for being used to cultivate hybridoma, in basis culture
Following components is contained on the basis of base：P-methylbenzoic acid, lipoic acid, linoleic acid, putrescine, monoethanolamine, zinc sulfate, gluathione
Peptide.
Further, the culture medium contains following components on the basis of basal medium：0.1-1mg/L is to methyl
Benzoic acid, 0.1-0.5mg/L lipoic acids, 0.01-0.1mg/L linoleic acid, 0.1-0.5mg/L putrescine, 1-5mg/L monoethanolamines,
0.1-0.5mg/L zinc sulfate, 1-5mg/L glutathione.
In a preferred embodiment, the basal medium is IMDM culture mediums.
In a preferred embodiment, the hybridoma is SP210 cell lines and immune mouse spleen B cell
Fusion is obtained.
Present invention also offers application of the above-mentioned culture medium in culture hybridoma.
Present invention also offers a kind of method for cultivating hybridoma, it is including the use of above-mentioned medium culture hybridoma
The step of cell.
Present invention also offers a kind of Hybridoma Cell Culture thing, it is obtained by above method culture.
The Growth of Hybridoma Cell rule of the medium culture of the present invention, compared with traditional Hybridoma Cell Culture base,
The hybridoma that culture is obtained in same time under the culture medium equal conditions of the present invention is more, and cell volume is bigger, cell
Form is fuller, and the duration cultivated is longer, and cell viability is higher, can be preferably applied to the production of monoclonal antibody.
Brief description of the drawings
Fig. 1 is thin for the sp210 and mouse spleen B lymphs respectively using embodiment 1-4 and traditional IMDM medium cultures
The microphoto of the obtained hybridoma of born of the same parents' fusion.
Embodiment
The principle and feature of the present invention are described below in conjunction with example, the given examples are served only to explain the present invention, and
It is non-to be used to limit the scope of the present invention.
1. the preparation of Hybridoma Cell Culture base
The culture medium of the present invention is obtained by the way that the composition shown in table 1 to be added to prepare in IMDM culture mediums.Table 1 is to IMDM
The composition added in culture medium
2. the culture of hybridoma
The hybridoma used in confirmatory experiment is the obtained hybridization that sp210 is merged with mouse spleen bone-marrow-derived lymphocyte
Oncocyte, the medium culture hybridoma obtained respectively with being prepared in above-described embodiment, using traditional IMDM culture mediums as
Control.
3. microscope inspection observation checking after cell attachment
In the case of initial density identical, by the hybridoma culture medium in embodiment 1-4 and traditional IMDM
Culture medium is under equal conditions cultivated, and is as a result shown, with times of the hybridoma of the medium culture in embodiment 1-4
The increasing time it is more traditional IMDM culture mediums it is shorter.The hybridoma that above-mentioned culture is obtained is put to be observed under the microscope, monoclonal antibody
The hybridoma of special culture media culture is relatively bigger (as shown in Figure 1), and vigor is higher.And in continuous culture more than 8 days,
The hybridoma of monoclonal antibody special culture media culture still can be with normal growth, and the hybridoma of conventional medium culture occurs
Broken, situation is deteriorated, it is seen that the continuation of the medium culture in above example is longer.
The foregoing is only presently preferred embodiments of the present invention, be not intended to limit the invention, it is all the present invention spirit and
Within principle, any modification, equivalent substitution and improvements made etc. should be included in the scope of the protection.
Claims (7)
1. a kind of be used to cultivate the culture medium of hybridoma, it is characterised in that contained on the basis of basal medium with
Lower component：P-methylbenzoic acid, lipoic acid, linoleic acid, putrescine, monoethanolamine, zinc sulfate, glutathione.
2. culture medium according to claim 1, it is characterised in that contained on the basis of basal medium with the following group
Point：0.1-1mg/L p-methylbenzoic acids, 0.1-0.5mg/L lipoic acids, 0.01-0.1mg/L linoleic acid, 0.1-0.5mg/L are rotten
Amine, 1-5mg/L monoethanolamines, 0.1-0.5mg/L zinc sulfate, 1-5mg/L glutathione.
3. culture medium according to claim 1, it is characterised in that the basal medium is IMDM culture mediums.
4. the culture medium according to any one of claim 1-3, it is characterised in that the hybridoma is that SP210 is thin
Born of the same parents are to merge to obtain with immune mouse spleen B cell.
5. application of the culture medium in culture hybridoma any one of claim 1-4.
6. a kind of method for cultivating hybridoma, it is characterised in that including the use of the training any one of claim 1-4
The step of supporting base culture hybridoma.
7. a kind of Hybridoma Cell Culture thing, it is characterised in that obtained by the method described in claim 6.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110297085A (en) * | 2019-06-13 | 2019-10-01 | 武汉博士德生物工程有限公司 | It is a kind of for enhancing the antibody diluent of immune-blotting method signal |
| CN111109248A (en) * | 2019-12-24 | 2020-05-08 | 武汉博士德生物工程有限公司 | Hybridoma cell cryopreservation liquid and 96-well plate cryopreservation recovery method |
| CN111808825A (en) * | 2020-09-14 | 2020-10-23 | 上海奥浦迈生物科技有限公司 | Selective medium for screening hybridoma cells and application |
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| CN1229851A (en) * | 1999-03-02 | 1999-09-29 | 华东理工大学 | Hybridizing tumour cell non-serum culture medium |
| CN101195817A (en) * | 2007-12-28 | 2008-06-11 | 天津百若克医药生物技术有限责任公司 | Hybrid tumor cell amplification culture medium and uses thereof |
| CN101418330A (en) * | 2008-06-20 | 2009-04-29 | 华东理工大学 | Non protein culture medium adapted to large-scale culture of NSO cell and production of antibody |
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| CN106190985A (en) * | 2015-04-29 | 2016-12-07 | 李昂 | Hybridizing tumour cell non-serum culture fluid |
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| CN1229851A (en) * | 1999-03-02 | 1999-09-29 | 华东理工大学 | Hybridizing tumour cell non-serum culture medium |
| CN101195817A (en) * | 2007-12-28 | 2008-06-11 | 天津百若克医药生物技术有限责任公司 | Hybrid tumor cell amplification culture medium and uses thereof |
| CN101418330A (en) * | 2008-06-20 | 2009-04-29 | 华东理工大学 | Non protein culture medium adapted to large-scale culture of NSO cell and production of antibody |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110297085A (en) * | 2019-06-13 | 2019-10-01 | 武汉博士德生物工程有限公司 | It is a kind of for enhancing the antibody diluent of immune-blotting method signal |
| CN111109248A (en) * | 2019-12-24 | 2020-05-08 | 武汉博士德生物工程有限公司 | Hybridoma cell cryopreservation liquid and 96-well plate cryopreservation recovery method |
| CN111808825A (en) * | 2020-09-14 | 2020-10-23 | 上海奥浦迈生物科技有限公司 | Selective medium for screening hybridoma cells and application |
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Application publication date: 20170808 |